Development of Simplified in Vitro P-Glycoprotein Substrate Assay and in Silico Prediction Models To Evaluate Transport Potential of P-Glycoprotein
| Autor: | Tomomi Taniguchi, Nao Torimoto-Katori, Tomoko Watanabe, Tsuyoshi Esaki, Kenji Mizuguchi, Tsuyoshi Takahashi, Reiko Watanabe, Yuko Ogasawara, Rikiya Ohashi, Mikiko Tsukimoto |
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| Rok vydání: | 2019 |
| Předmět: |
ATP Binding Cassette Transporter
Subfamily B Cell Membrane Permeability Swine In silico Drug Evaluation Preclinical Pharmaceutical Science Biological Availability 02 engineering and technology Transfection 030226 pharmacology & pharmacy Intestinal absorption Machine Learning 03 medical and health sciences 0302 clinical medicine In vivo Drug Discovery Animals Computer Simulation ATP Binding Cassette Transporter Subfamily B Member 1 P-glycoprotein biology Chemistry Drug discovery Substrate (chemistry) Reproducibility of Results 021001 nanoscience & nanotechnology Data Accuracy Protein Transport Intestinal Absorption Test set biology.protein Molecular Medicine LLC-PK1 Cells 0210 nano-technology Biological system Flux (metabolism) Central Nervous System Agents |
| Zdroj: | Molecular pharmaceutics. 16(5) |
| ISSN: | 1543-8392 |
| Popis: | For efficient drug discovery and screening, it is necessary to simplify P-glycoprotein (P-gp) substrate assays and to provide in silico models that predict the transport potential of P-gp. In this study, we developed a simplified in vitro screening method to evaluate P-gp substrates by unidirectional membrane transport in P-gp-overexpressing cells. The unidirectional flux ratio positively correlated with parameters of the conventional bidirectional P-gp substrate assay ( R2 = 0.941) and in vivo Kp,brain ratio (mdr1a/1b KO/WT) in mice ( R2 = 0.800). Our in vitro P-gp substrate assay had high reproducibility and required approximately half the labor of the conventional method. We also constructed regression models to predict the value of P-gp-mediated flux and three-class classification models to predict P-gp substrate potential (low-, medium-, and high-potential) using 2397 data entries with the largest data set collected under the same experimental conditions. Most compounds in the test set fell within two- and three-fold errors in the random forest regression model (71.3 and 88.5%, respectively). Furthermore, the random forest three-class classification model showed a high balanced accuracy of 0.821 and precision of 0.761 for the low-potential classes in the test set. We concluded that the simplified in vitro P-gp substrate assay was suitable for compound screening in the early stages of drug discovery and that the in silico regression model and three-class classification model using only chemical structure information could identify the transport potential of compounds including P-gp-mediated flux ratios. Our proposed method is expected to be a practical tool to optimize effective central nervous system (CNS) drugs, to avoid CNS side effects, and to improve intestinal absorption. |
| Databáze: | OpenAIRE |
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